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目的探讨交通相关PM2.5对Jurkat T细胞白介素-2(IL-2)的影响及钙信号通路对IL-2释放的调控作用。方法以100μg/ml的PM2.5染毒Jurkat T细胞,以生理盐水为对照组,并用钙调磷酸酶拮抗剂环孢菌素A(CSA)、钙离子螯合剂(EGTA)进行干预,设置空白滤膜组、EGTA组和CSA组平行对照,染毒时间为3、6和24 h。ELISA方法测定细胞上清IL-2水平;QRT-PCR测定细胞钙调磷酸酶(CaN)、活化T细胞转录因子(NFAT)的mRNA表达;免疫荧光法观察NFAT核分布情况。结果交通相关PM2.5染毒细胞3、6和24 h后,100μg/ml PM2.5组IL-2水平明显低于空白滤膜和生理盐水组,但高于100μg/ml PM2.5+CSA组、100μg/ml PM2.5+EGTA组,差异均有统计学意义(P<0.05),并且随着时间的延长,IL-2表达水平呈降低趋势。100μg/ml PM2.5组的NFAT、CaN mRNA表达水平高于对照组、100μg/ml PM2.5+CSA组、100μg/ml PM2.5+EGTA组,差异均具有统计学意义(P<0.05)。PM2.5能够使NFAT蛋白脱磷酸化而活化,可移位至细胞核内。白介素-2表达水平与NFAT基因、CaN基因表达水平呈负相关,差异均具有统计学意义(P<0.05)。结论交通相关PM2.5可抑制Jurkat T细胞释放IL-2,Ca2+-CaN-NFAT信号通路可能参与调控PM2.5对Jurkat T细胞IL-2的释放。
Objective To investigate the effects of traffic-related PM2.5 on interleukin-2 (IL-2) in Jurkat T cells and the regulatory effect of calcium signaling pathway on IL-2 release. Methods Jurkat T cells were exposed to PM2.5 at 100 μg / ml. Normal saline was used as a control and interventions were performed with calcineurin antagonists cyclosporine A (CSA) and calcium chelator (EGTA) The filter group, EGTA group and CSA group parallel control, exposure time was 3,6 and 24 h. ELISA method was used to determine the level of IL-2 in cell supernatant. The mRNA expression of calcineurin (CaN) and activated T cell transcription factor (NFAT) was detected by QRT-PCR. The NFAT nuclear distribution was observed by immunofluorescence. Results The levels of IL-2 in 100μg / ml PM2.5 group were significantly lower than those in blank filter group and saline group at 3, 6 and 24 h after exposure to PM2.5, but higher than 100μg / ml PM2.5 + CSA Group, 100μg / ml PM2.5 + EGTA group, the difference was statistically significant (P <0.05), and with the extension of time, IL-2 expression levels showed a downward trend. The levels of NFAT and CaN mRNA in 100μg / ml PM2.5 group were significantly higher than those in control group, 100μg / ml PM2.5 + CSA group and 100μg / ml PM2.5 + EGTA group (P <0.05) . PM2.5 can make NFAT dephosphorylation and activation, can be translocated to the nucleus. The expression of interleukin-2 was negatively correlated with NFAT gene and CaN gene expression, the differences were statistically significant (P <0.05). CONCLUSION: Transport-related PM2.5 can inhibit the release of IL-2 from Jurkat T cells. Ca2 + -CaN-NFAT signaling pathway may be involved in the regulation of PM2.5 release of Jurkat T cells by IL-2.